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Rockland Immunochemicals collagen type iv
Collagen Type Iv, supplied by Rockland Immunochemicals, used in various techniques. Bioz Stars score: 93/100, based on 68 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/600+401+106s/COLLAGEN+TYPE+IV+ANTIBODY/pmc12057615-218-82-98
Average 93 stars, based on 68 article reviews

collagen type iv - by Bioz Stars, 2026-09

93/100 stars

Images

Incubation:

Article Title: Lysophosphatidic Acid Modulates TGF-β2-Induced Biological Phenotype in Human Conjunctival Fibroblasts
Article Snippet: .. Briefly, 4% paraformaldehyde-fixed 3D spheroids were incubated in 3% BSA in PBS for 3 h. After washing twice with PBS for 30 min each time, 3D spheroids were sequentially exposed to 1:200 dilution of primary antibodies, including an anti-human COL1 (#600-401-103-0.1, ROCKLAND Antibodies & Assays, Limerick, PA, USA), COL4 (#600-401-106S, ROCKLAND Antibodies & Assays, Limerick, PA, USA), COL6 (#009-001-108, ROCKLAND Antibodies & Assays, Limerick, PA, USA), and FN (#A-11, Santa Cruz Biotechnology, Inc., Dallas, TX, USA) rabbit antibody at 4 °C overnight. .. The 3D spheroids were then washed 3 times with PBS for 1 h each time and exposed to 1:1000 dilution of secondary antibodies, including a goat anti-rabbit IgG (488 nm, #A27034, Invitrogen, Waltham, MA, USA) with phalloidin (594 nm, 1:1000 dilution, #17466-45-4, Cayman Chemical, Ann Arbor, MI, USA) and DAPI (1:1000 dilution, #D523-10, DOJINDO, Osaka, Japan) for 3 h. Then, after mounting on ProLong Gold Antifade Mountant with a cover glass, immunofluorescent images were taken with a Nikon A1 confocal microscope using a ×20 air objective at a resolution of 1024 × 1024 pixels.

Article Title: Cytoplasmic retention of the DNA/RNA-binding protein FUS ameliorates organ fibrosis in mice
Article Snippet: .. Membranes were incubated with the following primary antibodies: rabbit anti-FUS/TLS (4885, Cell Signaling Technology), rabbit anti–histone H3 (9715, Cell Signaling Technology), mouse anti–α-tubulin (3873, Cell Signaling Technology), rat anti–collagen IV α2NC1 (7071, Chondrex), rabbit anti–collagen IV (600-401-106, Rockland), rabbit anti–collagen I (66948, Cell Signaling Technology), rabbit anti-desmin (ab15200, Abcam), mouse anti-TIMP1 (sc-s21734, Santa Cruz Biotechnology). .. IRDye fluorescent dyes (LI-COR Inc.) were used as secondary antibodies, and membranes were processed through a LI-COR Odyssey infrared imaging system.

Staining:

Article Title: Cytoplasmic retention of the DNA/RNA-binding protein FUS ameliorates organ fibrosis in mice
Article Snippet: .. Paraffin sections were stained with mouse anti-FUS (sc-47711, Santa Cruz Biotechnology), rabbit anti-GFP (NB600-308, Novus Biological), rabbit anti–phospho-EGFR (Tyr1173; 4407, Cell Signaling Technology), or rabbit anti–collagen IV (600401106, Rockland) antibodies followed by Alexa Fluor 488 anti-mouse (A32723, Alexa Fluor) and Alexa Fluor 555 anti-rabbit (A21428, Alexa Fluor). .. Slides were then mounted using ProLong Gold Antifade Mountant with DAPI (P36931, Thermo Fisher Scientific).

Recombinant:

Article Title: Engineering scalable vascularized kidney organoids for in vivo glomerular filtration with human endothelial integration.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Anti-Actin, α-Smooth Muscle - Cy3 conjugated antibody Sigma-Aldrich C6198, RRID: AB_476856 Beta III TUBULIN Millipore MAB1637, RRID: AB_2210524 CD146 Abcam ab75769, RRID: AB_2143375 CD31 Abcam ab9498, RRID: AB_307284 CDH1 Abcam ab11512, RRID: AB_298118 Collagen IV α5 (B51 and H53 clones) Shigei Medical Research Institute SGE-C451 (B51), RRID: AB_2924380 and SGE-C453 (H53) Collagen IV α1 Rockland 600-401-106S, RRID: AB_11183330 EHD3 Novus Biologicals 31896 Integrin alpha2 (EPR5788) GeneTex GTX63576 LTL Vector lab B-1325, RRID: AB_2336558 LTL-Alexa Fluor 647 Bioworld 21511594 MECA-32 BD Biosciences 553849 MEIS1 Active motif 39796 MEIS1/2/3 Santa cruz sc-101850, RRID: AB_2143143 NEPHRIN (NPHS1) PROGEN GP-N2, RRID: AB_2904121 PAX2 Biolegend 901001, RRID: AB_2565001 PDGFR-β R&D systems AF385, RRID: AB_355339 NPHS2 Abcam ab50339, RRID: AB_882097 PODXL R&D systems AF1658, RRID: AB_354920 SALL1 R&D systems PP-K9814-00, RRID: AB_1964373 SIX2 Proteintech 11562-1-AP, RRID: AB_2189084 WT1 Santa cruz sc-192, RRID: AB_632611 Chemicals, peptides, recombinant proteins, consumables StemFit® Basic02 Amsbio SFB-500 StemFit® Basic04 complete Amsbio SFB-504-CT LDEV-Free hESC-qualified Geltrex Life Technologies A1413302 AccutaseTM STEMCELL TECHNOLOGIES 07920 PBS Life Technologies 10010-023 24-well tissue culture plates TPP 92024 Y27632 TOCRIS 1254 FGF2 Peprotech 100-18B aRPMI1640 Life Technologies 12633-020 L-GlutaMAX Life Technologies 35050-061 CHIR99021 TOCRIS 4423 Dorsomorphin TOCRIS 3093 Activin R&D 338-AC-050 96-well, round-bottom, ultra-low attachment plates Corning 7007 Fig. 6 | Implantation of pre-vascularized human nephron sheets into mice facilitates size-selective filtration of LMW dextran via graft-host anastomosis of human andmurine capillaries. ..

Filtration:

Bioprocessing:

Article Title: Functionalized Cerium Oxide Nanoparticles Enhance Penetration into Melanoma Spheroids In Vivo through Angiogenesis
Article Snippet: .. Cells were blocked with BSA (1% w/v) in PBS for 16 h at 4 °C and then washed twice with PBS containing Tween‐20 (PBS‐T; 0.1% w/v) before the addition of either Rhodamine‐phalloidin (1:200 in PBS‐T; Thermo Fisher Scientific) at RT for 1 h in the dark and DAPI (1:1000, Thermo Fisher Scientific) for 30 min at RT or primary antibodies at RT for 2 h. Primary antibodies included mouse monoclonal antibodies against perlecan (2 mg mL −1 ; clone A74; AbCam) or collagen type IV (2 mg mL −1 ; clone Col94) or a rabbit polyclonal anti‐laminin antibody (Rockland). .. Cells were then incubated with either goat anti‐mouse Alexa Fluor 488 (1:500; Invitrogen) or goat anti‐rabbit Alexa Fluor 488 (1:500; Invitrogen) at RT for 1 h in the dark and counterstained with CellMask Deep red membrane stain (1:1000; Invitrogen) at RT for 1 h. Samples were washed twice with PBS‐T and a final rinse with PBS at 4 °C prior to imaging using a confocal microscope (IX70, Olympus or SP8, Leica).

Image Search Results

A Schematic procedures for kidney organoid differentiation from human PSC lines in static and STR culture systems. The schema explains the maintenance and differentiation process starting from PSCs to nephron progenitor cells (NPCs) and ultimately kidney organoids. STR and static organoids were treated with Activin for 2 or 3 days after 4 days of CHIR ± Dorsomorphin treatment. Spheroids were then treated with both CHIR and FGF9 starting on day 7 or 8 for 2 days, and subsequently, CHIR was removed. From day 13 or 14, no growth factors were added. The right-sided photo compares the final volumes of static (from 6 wells of one plate) and STR organoids, respectively. The scale bars: 25 mm. B Brightfield image of differentiating H9 cells in static and STR systems. The scale bar represents 500 µm. C Confocal images for the expression of nephron progenitor markers (SIX2 and SALL1) by H9 NPCs on day 8 of differentiation in static and STR systems. Scale bars: 100 µm. D Confocal images of day 21 STR-kidney organoids composed of proximal tubules (LTL: Lotus tetragonolobus lectin + ), interstitial cells (MEIS1/2/3 + ), endothelial cells (CD31 + ), podocytes (PODXL: Podocalyxin + ), loops of Henle/distal nephrons (CDH1: E-cadherin + ) with a scale bar of 200 µm. E Comparison of the total cost of goods/materials (COGs; cell culture reagents, media, and materials) in static and STR systems to generate the same cell number of kidney organoids. F Organoid cell numbers per media volume generated from one 96-well plate (static) and one 5-ml bioreactor (STR).

Journal: Npj Biomedical Innovations

Article Title: Engineering scalable vascularized kidney organoids for in vivo glomerular filtration with human endothelial integration

doi: 10.1038/s44385-025-00063-5

Figure Lengend Snippet: A Schematic procedures for kidney organoid differentiation from human PSC lines in static and STR culture systems. The schema explains the maintenance and differentiation process starting from PSCs to nephron progenitor cells (NPCs) and ultimately kidney organoids. STR and static organoids were treated with Activin for 2 or 3 days after 4 days of CHIR ± Dorsomorphin treatment. Spheroids were then treated with both CHIR and FGF9 starting on day 7 or 8 for 2 days, and subsequently, CHIR was removed. From day 13 or 14, no growth factors were added. The right-sided photo compares the final volumes of static (from 6 wells of one plate) and STR organoids, respectively. The scale bars: 25 mm. B Brightfield image of differentiating H9 cells in static and STR systems. The scale bar represents 500 µm. C Confocal images for the expression of nephron progenitor markers (SIX2 and SALL1) by H9 NPCs on day 8 of differentiation in static and STR systems. Scale bars: 100 µm. D Confocal images of day 21 STR-kidney organoids composed of proximal tubules (LTL: Lotus tetragonolobus lectin + ), interstitial cells (MEIS1/2/3 + ), endothelial cells (CD31 + ), podocytes (PODXL: Podocalyxin + ), loops of Henle/distal nephrons (CDH1: E-cadherin + ) with a scale bar of 200 µm. E Comparison of the total cost of goods/materials (COGs; cell culture reagents, media, and materials) in static and STR systems to generate the same cell number of kidney organoids. F Organoid cell numbers per media volume generated from one 96-well plate (static) and one 5-ml bioreactor (STR).

Article Snippet: Collagen IV α1 , Rockland , 600-401-106S, RRID: AB_11183330.

Techniques: Expressing, Comparison, Cell Culture, Generated

A-D Principal component ( A ) and hierarchical clustering ( B ) analyses from RNA-seq samples on day 0, day 8 (NPC), static day 21, 35, 49, 63 (Org_21, Org_35, Org_49, Org_63), and STR day 21, 35, 49 (STR_21, STR_35, STR_49). n = 3 mRNA samples. Each mRNA sample was collected from 6-12 organoids. Gene ontology (GO) analysis of DEGs significantly upregulated in STR organoids compared to static ones on day 21 ( C ) and day 35 ( D ). E Live-cell imaging in STR and static organoids to assess the transepithelial cation transport using Rhodamine 123 (Rh123). On days 32–38 of differentiation, both STR and static ones were transferred to the 1% Geltrex-coated 8-well glass-bottom chamber slides. On days 35–39, proximal tubules of organoids in these slides were labeled with LTL conjugated with Alexa Fluor 647. On days 36–40, after the organoids in chamber slides were assigned and mapped by the confocal microscope, the cation Rh123 (10 mM) was added into the culture media at a final concentration of 10 μM. Live-cell imaging was immediately started to check the efficiency of cation transport through LTL+ proximal tubular epithelial cells. Baseline brightfield images show the particular LTL+ proximal tubules before the addition of Rh123. Following the supplementation of Rh123, its uptake by tubular epithelial cells was recorded for 20–30 min via real-time imaging. The images encircled by white dashed lines exhibit the course of Rh123 uptake by LTL+ tubular epithelial cells at 0, 10, and 20 min. F Quantification of apical and basal Rh123 fluorescence intensities of proximal tubular epithelial cells to calculate the apicobasal ratio of Rh123 uptake for the assessment of cation transport from basal to apical sides in these tubular epithelial cells. Each dot represents the proximal tubular histogram analysis of each organoid, n = 6 organoids from 2 independent organoid batches of BJFF.6 and HUES62 cell lines. The asterisk in the bar graph indicates the p value derived from a two-tailed unpaired t-test. Means ± SEM. ** p ≤ 0.01.

Journal: Npj Biomedical Innovations

Article Title: Engineering scalable vascularized kidney organoids for in vivo glomerular filtration with human endothelial integration

doi: 10.1038/s44385-025-00063-5

Figure Lengend Snippet: A-D Principal component ( A ) and hierarchical clustering ( B ) analyses from RNA-seq samples on day 0, day 8 (NPC), static day 21, 35, 49, 63 (Org_21, Org_35, Org_49, Org_63), and STR day 21, 35, 49 (STR_21, STR_35, STR_49). n = 3 mRNA samples. Each mRNA sample was collected from 6-12 organoids. Gene ontology (GO) analysis of DEGs significantly upregulated in STR organoids compared to static ones on day 21 ( C ) and day 35 ( D ). E Live-cell imaging in STR and static organoids to assess the transepithelial cation transport using Rhodamine 123 (Rh123). On days 32–38 of differentiation, both STR and static ones were transferred to the 1% Geltrex-coated 8-well glass-bottom chamber slides. On days 35–39, proximal tubules of organoids in these slides were labeled with LTL conjugated with Alexa Fluor 647. On days 36–40, after the organoids in chamber slides were assigned and mapped by the confocal microscope, the cation Rh123 (10 mM) was added into the culture media at a final concentration of 10 μM. Live-cell imaging was immediately started to check the efficiency of cation transport through LTL+ proximal tubular epithelial cells. Baseline brightfield images show the particular LTL+ proximal tubules before the addition of Rh123. Following the supplementation of Rh123, its uptake by tubular epithelial cells was recorded for 20–30 min via real-time imaging. The images encircled by white dashed lines exhibit the course of Rh123 uptake by LTL+ tubular epithelial cells at 0, 10, and 20 min. F Quantification of apical and basal Rh123 fluorescence intensities of proximal tubular epithelial cells to calculate the apicobasal ratio of Rh123 uptake for the assessment of cation transport from basal to apical sides in these tubular epithelial cells. Each dot represents the proximal tubular histogram analysis of each organoid, n = 6 organoids from 2 independent organoid batches of BJFF.6 and HUES62 cell lines. The asterisk in the bar graph indicates the p value derived from a two-tailed unpaired t-test. Means ± SEM. ** p ≤ 0.01.

Article Snippet: Collagen IV α1 , Rockland , 600-401-106S, RRID: AB_11183330.

Techniques: RNA Sequencing, Live Cell Imaging, Labeling, Microscopy, Concentration Assay, Imaging, Fluorescence, Derivative Assay, Two Tailed Test

A Vessel sprouts in polarized podocyte clusters with CD146+ capillary network supported by MEIS1/2/3+ stromal cells in day 21 STR organoids (left set of two images) compared to the podocyte clusters of day 21 control static organoids (right set of four images). White arrows in the STR images indicate the vascularized glomeruli surrounded by polarized podocytes. Scale bars of STR images represent 100 μm and 10 μm (zoomed STR image), respectively. Rectangular white dashed lines point out the zoomed images of non-vascularized podocyte clusters of the static organoid. Scale bars of static organoid images represent 100 μm. B 3D surface reconstruction of CD146+ capillary network and surrounding PODXL+ podocyte clusters as vascularized glomerular components. Z-stack images were acquired at a step size of 2 μm. The scale bar represents 30 μm. C Transmission electron microscopy images show multiple vascular lumens (left image) and accompanying podocyte foot processes around the vascular lumen (right image) within organoid glomerular structures on day 35. The asterisk indicates the nucleus of the podocyte surrounding the vascular lumen, while the arrowheads mark podocyte foot processes. Black scale bars at the corners: 2 μm. D Percentages of vascularized podocyte clusters containing CD146 endothelial surface in STR versus static organoids via 3D surface reconstruction by machine learning. n = 3 static and n = 9 STR organoids from 3 independent batches differentiated from BJFF.6 or HUES62 cells. E ACTA2+ mesangial-like cells surrounded by polarized PODXL+ podocytes in Bowman’s space of day 35 STR and static organoids. STR organoids were generated by the transfer of the same-batch static organoids into STR wells on day 14 of differentiation. The graph represents the percentage of organoids containing ACTA2+ cells ( n = 5 static and n = 6 STR organoids from 2 independent batches differentiated from H9 or HUES62 cells). F Upregulation of certain GO terms associated with extracellular matrix organization, cell adhesion, and migration in day 35 STR organoids compared to static organoids. “Regulation of cell migration”, “Positive regulation of cell adhesion”, “Regulation of cell adhesion”, and “Extracellular matrix organization” terms constitute GO biological processes while “Extracellular matrix”, “Collagen-containing extracellular matrix”, “Actin-based cell projection”, and “Basement membrane”, “Collagen network”, and “Basement membrane collagen trimer” terms can be classified as GO cellular components, and “Assembly of collagen fibrils and other multimeric structures” is a part of Reactome pathways. G Inhibition of glomerular vascularization of STR organoids by the application of α2β1 integrin inhibitor, BTT3033 (stock concentration of 10 mM), with a final concentration of 1 μM for 7 or 18 days, in contrast to the control vehicle-treated organoids. The white arrows in the confocal microscopy images indicate the vascularized glomeruli surrounded by polarized podocytes. The scale bars represent 100 μm. H 3D surface reconstruction of CD146+ capillary network and surrounding PODXL+ podocyte clusters as vascularized glomerular surfaces in vehicle- and BTT3033-treated organoids. Z-stack images were acquired at a step size of 2 μm. The scale bar represents 50 μm. Y-axes of graphs represent two parameters: the percentage of vascularized podocyte clusters and the volume percent ratio of CD146+ vascular surface in these podocyte clusters to the whole PODXL+ podocyte surface, respectively ( n = 7). Asterisks in the bar graphs ( D , E , H ) indicate p values derived from two-tailed unpaired t-tests. Means ± SEM. (* p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001).

Journal: Npj Biomedical Innovations

Article Title: Engineering scalable vascularized kidney organoids for in vivo glomerular filtration with human endothelial integration

doi: 10.1038/s44385-025-00063-5

Figure Lengend Snippet: A Vessel sprouts in polarized podocyte clusters with CD146+ capillary network supported by MEIS1/2/3+ stromal cells in day 21 STR organoids (left set of two images) compared to the podocyte clusters of day 21 control static organoids (right set of four images). White arrows in the STR images indicate the vascularized glomeruli surrounded by polarized podocytes. Scale bars of STR images represent 100 μm and 10 μm (zoomed STR image), respectively. Rectangular white dashed lines point out the zoomed images of non-vascularized podocyte clusters of the static organoid. Scale bars of static organoid images represent 100 μm. B 3D surface reconstruction of CD146+ capillary network and surrounding PODXL+ podocyte clusters as vascularized glomerular components. Z-stack images were acquired at a step size of 2 μm. The scale bar represents 30 μm. C Transmission electron microscopy images show multiple vascular lumens (left image) and accompanying podocyte foot processes around the vascular lumen (right image) within organoid glomerular structures on day 35. The asterisk indicates the nucleus of the podocyte surrounding the vascular lumen, while the arrowheads mark podocyte foot processes. Black scale bars at the corners: 2 μm. D Percentages of vascularized podocyte clusters containing CD146 endothelial surface in STR versus static organoids via 3D surface reconstruction by machine learning. n = 3 static and n = 9 STR organoids from 3 independent batches differentiated from BJFF.6 or HUES62 cells. E ACTA2+ mesangial-like cells surrounded by polarized PODXL+ podocytes in Bowman’s space of day 35 STR and static organoids. STR organoids were generated by the transfer of the same-batch static organoids into STR wells on day 14 of differentiation. The graph represents the percentage of organoids containing ACTA2+ cells ( n = 5 static and n = 6 STR organoids from 2 independent batches differentiated from H9 or HUES62 cells). F Upregulation of certain GO terms associated with extracellular matrix organization, cell adhesion, and migration in day 35 STR organoids compared to static organoids. “Regulation of cell migration”, “Positive regulation of cell adhesion”, “Regulation of cell adhesion”, and “Extracellular matrix organization” terms constitute GO biological processes while “Extracellular matrix”, “Collagen-containing extracellular matrix”, “Actin-based cell projection”, and “Basement membrane”, “Collagen network”, and “Basement membrane collagen trimer” terms can be classified as GO cellular components, and “Assembly of collagen fibrils and other multimeric structures” is a part of Reactome pathways. G Inhibition of glomerular vascularization of STR organoids by the application of α2β1 integrin inhibitor, BTT3033 (stock concentration of 10 mM), with a final concentration of 1 μM for 7 or 18 days, in contrast to the control vehicle-treated organoids. The white arrows in the confocal microscopy images indicate the vascularized glomeruli surrounded by polarized podocytes. The scale bars represent 100 μm. H 3D surface reconstruction of CD146+ capillary network and surrounding PODXL+ podocyte clusters as vascularized glomerular surfaces in vehicle- and BTT3033-treated organoids. Z-stack images were acquired at a step size of 2 μm. The scale bar represents 50 μm. Y-axes of graphs represent two parameters: the percentage of vascularized podocyte clusters and the volume percent ratio of CD146+ vascular surface in these podocyte clusters to the whole PODXL+ podocyte surface, respectively ( n = 7). Asterisks in the bar graphs ( D , E , H ) indicate p values derived from two-tailed unpaired t-tests. Means ± SEM. (* p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001).

Article Snippet: Collagen IV α1 , Rockland , 600-401-106S, RRID: AB_11183330.

Techniques: Control, Transmission Assay, Electron Microscopy, Generated, Migration, Membrane, Inhibition, Concentration Assay, Confocal Microscopy, Derivative Assay, Two Tailed Test

A Workflow for the implantation of nephron sheets generated from STR organoids into dorsal skinfold chambers (DSFCs) of NSG mice. Implantation of STR organoids into mice was performed six times from three independent organoid batches. B Phase 1 left photo shows the self-assembled nephron sheet formed by STR organoids. Instead of implantation, this sheet was fixed with 4% PFA on day 28 to confirm the self-assembly process. The right image represents the stained nephron sheet with the designated markers (LTL, CD31, CDH1, CD146, PODXL, MEIS1/2/3) following the fixation process. The scale bars represent 1 mm. C Phase 2 photos include the NSG mouse with a dorsal skin fold chamber (DSFC) on its back (left photo) and the implanted nephron sheet generated from day 24 STR organoids inside the DSFC (right photo). D Phase 3 multiphoton intravital microscopy (MP-IVM) of the implanted sheet reveals the nephron units (designated by a circular white dashed line) with the dynamic flow of both high-molecular-weight (HMW, 500 kDa-Cy5) and low-molecular-weight (LMW, 3 kDa-FITC) dextrans in glomerular vascular-like structures (designated by 3D-reconstructed gray surface), accompanied by the filtration of LMW dextran into the surrounding space. The 3D-reconstructed cellular surface was generated via 3D surface rendering of Hoechst in Z-stack images by IMARIS. The scale bar displays 20 μm. E Phase 4 left photo shows the gross vascularization of the implanted nephron sheet with murine vessels on the 6th day of implantation, following its extraction and fixation. 3D reconstructed image (right) demonstrates the ongoing CD146+ human vascular network inside the same phase 4 nephron sheet (PODXL, CD146, LTL, MEIS123). Z-stack images were acquired at a step size of 100 μm. The scale bars represent 1 mm. F The confocal microscopy image showing the anastomosis of MECA-32-CD146+ human (white arrows, single-positive red) and MECA-32+ murine vascular endothelial networks. The scale bar displays 150 μm.

Journal: Npj Biomedical Innovations

Article Title: Engineering scalable vascularized kidney organoids for in vivo glomerular filtration with human endothelial integration

doi: 10.1038/s44385-025-00063-5

Figure Lengend Snippet: A Workflow for the implantation of nephron sheets generated from STR organoids into dorsal skinfold chambers (DSFCs) of NSG mice. Implantation of STR organoids into mice was performed six times from three independent organoid batches. B Phase 1 left photo shows the self-assembled nephron sheet formed by STR organoids. Instead of implantation, this sheet was fixed with 4% PFA on day 28 to confirm the self-assembly process. The right image represents the stained nephron sheet with the designated markers (LTL, CD31, CDH1, CD146, PODXL, MEIS1/2/3) following the fixation process. The scale bars represent 1 mm. C Phase 2 photos include the NSG mouse with a dorsal skin fold chamber (DSFC) on its back (left photo) and the implanted nephron sheet generated from day 24 STR organoids inside the DSFC (right photo). D Phase 3 multiphoton intravital microscopy (MP-IVM) of the implanted sheet reveals the nephron units (designated by a circular white dashed line) with the dynamic flow of both high-molecular-weight (HMW, 500 kDa-Cy5) and low-molecular-weight (LMW, 3 kDa-FITC) dextrans in glomerular vascular-like structures (designated by 3D-reconstructed gray surface), accompanied by the filtration of LMW dextran into the surrounding space. The 3D-reconstructed cellular surface was generated via 3D surface rendering of Hoechst in Z-stack images by IMARIS. The scale bar displays 20 μm. E Phase 4 left photo shows the gross vascularization of the implanted nephron sheet with murine vessels on the 6th day of implantation, following its extraction and fixation. 3D reconstructed image (right) demonstrates the ongoing CD146+ human vascular network inside the same phase 4 nephron sheet (PODXL, CD146, LTL, MEIS123). Z-stack images were acquired at a step size of 100 μm. The scale bars represent 1 mm. F The confocal microscopy image showing the anastomosis of MECA-32-CD146+ human (white arrows, single-positive red) and MECA-32+ murine vascular endothelial networks. The scale bar displays 150 μm.

Article Snippet: Collagen IV α1 , Rockland , 600-401-106S, RRID: AB_11183330.

Techniques: Generated, Staining, Intravital Microscopy, High Molecular Weight, Molecular Weight, Filtration, Extraction, Confocal Microscopy

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collagen type iv (Rockland Immunochemicals)

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